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BackgroundAedes-borne disease risk is associated with contemporary urbanization practices where city developing structures function as a catalyst for creating mosquito breeding habitats. We lack better understanding on how the links between landscape ecology and urban geography contribute to the prevalence and abundance of mosquito and pathogen spread.MethodsAn outdoor longitudinal study in Bengaluru (Karnataka, India) was conducted between February 2021 and June 2022 to examine the effects of macrohabitat types on the diversity and distribution of larval habitats, mosquito species composition, and body size to quantify the risk of dengue outbreak in the landscape context.FindingsA total of 8,717 container breeding sites were inspected, of these 1,316 were wet breeding habitats. A total of 1,619 mosquito larvae representing 16 species from six macrohabitats and nine microhabitats were collected. Aedes aegypti and Aedes albopictus were the dominant species and significantly higher in artificial habitats than in natural habitats. Breeding preference ratio for Aedes species was high in grinding stones and storage containers. The Aedes infestation indices were higher than the WHO threshold and showed significant linear increase from Barren habitat to High density areas. We found Ae. albopictus breeding in sympatry with Ae. aegypti had shorter wing length.ConclusionsA large proportion of larval habitats were man-made artificial containers. Landscape ecology drives mosquito diversity and abundance even at a small spatial scale which could be affecting the localized outbreaks. Our findings showed that sampling strategies for mosquito surveillance must include urban environments with non-residential locations and dengue transmission reduction programmes should focus on 'neighbourhood surveillance' as well to prevent and control the rising threat of Aedes-borne diseases.

Abstract The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2, has devastated every sphere of human society. Governments around the world implemented unprecedented policies designed to slow the spread of the disease and assistance to cope with its impacts. Such policies, however, are short-term and debates have ensued about what broader policies are needed in the post-COVID-19 era to ensure societies are better prepared for future pandemics. Public opinion concerning COVID-19 and the post-COVID-19 era is diverse, and the patterns in opinion are not well documented. Here we synthesized the opinions of 3731 research scholars throughout the world based on a survey. The highest consensus among respondents concerned the need for improving public health infrastructure and delivering economic support, whereas agreement concerning ecological aspects was low. The survey revealed three dimensions of thinking about COVID-19. The first dimension relates to public health and has widespread support. The second dimension relates to science-led policy development focusing on social justice and environmental governance, covering components of both ecology and economy. The third dimension covers the role of nature conservation in reducing the risk of pandemics. Although opinions differed with age, country of citizenship, and level of education, there is strong agreement on the need for global health equity and science-led public policy.

Abstract Background Vivax malaria is associated with significant morbidity and economic loss, and constitutes the bulk of malaria cases in large parts of Asia and South America as well as recent case reports in Africa. The widespread prevalence of vivax is a challenge to global malaria elimination programmes. Vivax malaria control is particularly challenged by existence of dormant liver stage forms that are difficult to treat and are responsible for multiple relapses, growing drug resistance to the asexual blood stages and host-genetic factors that preclude use of specific drugs like primaquine capable of targeting Plasmodium vivax liver stages. Despite an obligatory liver-stage in the Plasmodium life cycle, both the difficulty in obtaining P. vivax sporozoites and the limited availability of robust host cell models permissive to P. vivax infection are responsible for the limited knowledge of hypnozoite formation biology and relapse mechanisms, as well as the limited capability to do drug screening. Although India accounts for about half of vivax malaria cases world-wide, very little is known about the vivax liver stage forms in the context of Indian clinical isolates. Methods To address this, methods were established to obtain infective P. vivax sporozoites from an endemic region in India and multiple assay platforms set up to detect and characterize vivax liver stage forms. Different hepatoma cell lines, including the widely used HCO4 cells, primary human hepatocytes as well as hepatocytes obtained from iPSC’s generated from vivax patients and healthy donors were tested for infectivity with P. vivax sporozoites. Results Both large and small forms of vivax liver stage are detected in these assays, although the infectivity obtained in these platforms are low. Conclusions This study provides a proof of concept for detecting liver stage P. vivax and provide the first characterization of P. vivax liver stage forms from an endemic region in India.

Background & objectives: In India, the burden of Plasmodium vivax malaria has been projected to be highest in some areas. This study investigated the efficacy and safety of fixed dose combination (FDC) of arterolane maleate (AM) 37.5 mg and piperaquine phosphate 187.5 mg (PQP) dispersible tablets and (not with) chloroquine in the treatment of uncomplicated vivax malaria in pediatric patients. Methods: This multicentric, open-label trial was carried out at 12 sites in India. A total of 164 patients aged 6 months to 12 years with P. vivax malaria were randomized in a ratio of 2:1 to AM-PQP (111 patients) or chloroquine (53 patients) arms. The duration of follow up was 42 days. Results: At 72 hours, the proportion of a parasitaemic and afebrile patients was 100% in both treatment arms in per protocol (PP) population, and 98.2% and 100% [95% CI: -1.8 (-6.33 to 5.08)] in AM-PQP and chloroquine arms, respectively, in intent to treat (ITT) population. The efficacy and safety of AM-PQP was found to be comparable to chloroquine in the treatment of uncomplicated P. vivax malaria in pediatric patients. Overall, the cure rate at Day 28 and 42 was >95% for both AM-PQP or CQ. The commonly reported clinical adverse event was vomiting. No patient was discontinued for any QTc abnormality. Interpretation & conclusion: The efficacy and safety of FDC of arterolane maleate and piperaquine phosphate was found to be comparable to chloroquine for treatment of uncomplicated P. vivax malaria in pediatric patients.

Background & objectives: Understanding of malaria vector distribution and influence of climatic environments is essential for devising control strategies. The aim of the study was to study the bionomics of prevalent malaria vectors in three different settings for development of evidence-based sustainable malaria control strategy with special reference to vector control. Methods: Three villages with different eco-epidemiological settings like riverine-low malarious, riverine-high malarious and non-riverine high malarious villages were selected after baseline studies. Entomological aspects such as man hour density, per structure density, mosquito landing collections, sibling species identification, insecticide susceptibility status, parity rate, etc. were studied in these three villages following standard methods and techniques. The effect of these variables was analysed statistically. Results: Mosquito collections revealed the presence of three malaria vectors in the study villages, namely Anopheles culicifacies s.l., An. fluviatilis s.l. and An. stephensi (Diptera: Culicidae) with varying proportions and seasonal abundance. Densities of the principal malaria vector, An. culicifacies varied seasonally. Anopheles culicifacies was found resistant to DDT (4%), malathion (5%), lambda-cyhalothrin (0.05%) and alpha-cypermethrin (0.1%). Peak density of An. culicifacies was found during post-monsoon months starting from August-September to December in the high malarious villages. Interpretation & conclusion: The main vector control interventions should be planned in the post-monsoon months in these villages and suitable insecticide resistance management strategy should be followed as An. culicifacies was found resistant to DDT, malathion, alpha-cypermethrin and lambda-cyhalothrin in the study area.

Abstract Background The native gut microbiota of Anopheles mosquitoes is known to play a key role in the physiological function of its host. Interestingly, this microbiota can also influence the development of Plasmodium in its host mosquitoes. In recent years, much interest has been shown in the employment of gut symbionts derived from vectors in the control of vector-borne disease transmission. In this study, the midgut microbial diversity has been characterized among laboratory-reared adult Anopheles stephensi mosquitoes, from the colony created by rearing progeny of wild-caught mosquitoes (obtained from three different locations in southern India) for multiple generations, using 16S ribosomal RNA (rRNA) gene sequencing approach. Further, the influence of native midgut microbiota of mosquitoes on the development of rodent malaria parasite Plasmodium berghei in its host has been studied. Methods The microbial diversity associated with the midgut of An. stephensi mosquitoes was studied by sequencing V3 region of 16S ribosomal RNA (rRNA) gene. The influence of native midgut microbiota of An. stephensi mosquitoes on the susceptibility of the mosquitoes to rodent malaria parasite P. berghei was studied by comparing the intensity and prevalence of P. berghei infection among the antibiotic treated and untreated cohorts of mosquitoes. Results The analysis of bacterial diversity from the midguts of An. stephensi showed Proteobacteria as the most dominant population among the three laboratory-reared strains of An. stephensi studied. Major genera identified among these mosquito strains were Acinetobacter, Pseudomonas, Prevotella, Corynebacterium, Veillonella, and Bacillus. The mosquito infectivity studies carried out to determine the implication of total midgut microbiota on P. berghei infection showed that mosquitoes whose native microbiota cleared with antibiotics had increased susceptibility to P. berghei infection compared to the antibiotic untreated mosquitoes with its natural native microbiota. Conclusions The use of microbial symbiont to reduce the competence of vectors involved in disease transmission has gained much importance in recent years as an emerging alternative approach towards disease control. In this context, the present study was aimed to identify the midgut microbiota composition of An. stephensi, and its effect on the development of P. berghei. Interestingly, the analysis of midgut microbiota from An. stephensi revealed the presence of genus Veillonella in Anopheles species for the first time. Importantly, the study also revealed the negative influence of total midgut microbiota on the development of P. berghei in three laboratory strains of An. stephensi, emphasizing the importance of understanding the gut microbiota in malaria vectors, and its relationship with parasite development in designing strategies to control malaria transmission.

Heme metabolism is central to malaria parasite biology. The parasite acquires heme from host hemoglobin in the intraerythrocytic stages and stores it as hemozoin to prevent free heme toxicity. The parasite can also synthesize heme de novo, and all the enzymes in the pathway are characterized. To study the role of the dual heme sources in malaria parasite growth and development, we knocked out the first enzyme, δ-aminolevulinate synthase (ALAS), and the last enzyme, ferrochelatase (FC), in the heme-biosynthetic pathway of Plasmodium berghei (Pb). The wild-type and knockout (KO) parasites had similar intraerythrocytic growth patterns in mice. We carried out in vitro radiolabeling of heme in Pb-infected mouse reticulocytes and Plasmodium falciparum-infected human RBCs using [4-(14)C] aminolevulinic acid (ALA). We found that the parasites incorporated both host hemoglobin-heme and parasite-synthesized heme into hemozoin and mitochondrial cytochromes. The similar fates of the two heme sources suggest that they may serve as backup mechanisms to provide heme in the intraerythrocytic stages. Nevertheless, the de novo pathway is absolutely essential for parasite development in the mosquito and liver stages. PbKO parasites formed drastically reduced oocysts and did not form sporozoites in the salivary glands. Oocyst production in PbALASKO parasites recovered when mosquitoes received an ALA supplement. PbALASKO sporozoites could infect mice only when the mice received an ALA supplement. Our results indicate the potential for new therapeutic interventions targeting the heme-biosynthetic pathway in the parasite during the mosquito and liver stages.

Recent reports highlight the severity and the morbidity of disease caused by the long neglected malaria parasite Plasmodium vivax. Due to inherent difficulties in the laboratory-propagation of P. vivax, the biology of this parasite has not been adequately explored. While the proteome of P. falciparum, the causative agent of cerebral malaria, has been extensively explored from several sources, there is limited information on the proteome of P. vivax. We have, for the first time, examined the proteome of P. vivax isolated directly from patients without adaptation to laboratory conditions. We have identified 153 proteins from clinical P. vivax, majority of which do not show homology to any previously known gene products. We also report 29 new proteins that were found to be expressed in P. vivax for the first time. In addition, several proteins previously implicated as anti-malarial targets, were also found in our analysis. Most importantly, we found several unique proteins expressed by P. vivax.This study is an important step in providing insight into physiology of the parasite under clinical settings.